PI3K mediated activation of GSK-3β reduces at-level primary afferent growth responses associated with excitotoxic spinal cord injury dysesthesias

Molecular Pain

Kori L Brewer²  Elizabeth Dugan¹  Sonja K Bareiss¹ 

[1] Department of Physical Therapy, College of Allied Health Sciences, East Carolina University, 600 Moye Blvd., Room 2405E, Greenville 27834, NC, USA;Department of Emergency Medicine, East Carolina University, 600 Moye Blvd., Room 2405E, Greenville 27834, NC, USA

关键词: Neurite outgrowth;    Dorsal root ganglia (DRG);    Dysesthesias;    Pain;    Spinal cord injury (SCI);    GSK-3β;   

Others  :  1217330
DOI  :  10.1186/s12990-015-0041-2

 received in 2015-03-13, accepted in 2015-06-11,  发布年份 2015

【 摘 要 】

Background

Neuropathic pain and sensory abnormalities are a debilitating secondary consequence of spinal cord injury (SCI). Maladaptive structural plasticity is gaining recognition for its role in contributing to the development of post SCI pain syndromes. We previously demonstrated that excitotoxic induced SCI dysesthesias are associated with enhanced dorsal root ganglia (DRG) neuronal outgrowth. Although glycogen synthase kinase-3β (GSK-3β) is a known intracellular regulator neuronal growth, the potential contribution to primary afferent growth responses following SCI are undefined. We hypothesized that SCI triggers inhibition of GSK-3β signaling resulting in enhanced DRG growth responses, and that PI3K mediated activation of GSK-3β can prevent this growth and the development of at-level pain syndromes.

Results

Excitotoxic SCI using intraspinal quisqualic acid (QUIS) resulted in inhibition of GSK-3β in the superficial spinal cord dorsal horn and adjacent DRG. Double immunofluorescent staining showed that GSK-3β ^Pwas expressed in DRG neurons, especially small nociceptive, CGRP and IB4-positive neurons. Intrathecal administration of a potent PI3-kinase inhibitor (LY294002), a known GSK-3β activator, significantly decreased GSK-3β ^Pexpression levels in the dorsal horn. QUIS injection resulted in early (3 days) and sustained (14 days) DRG neurite outgrowth of small and subsequently large fibers that was reduced with short term (3 days) administration of LY294002. Furthermore, LY294002 treatment initiated on the date of injury, prevented the development of overgrooming, a spontaneous at-level pain related dysesthesia.

Conclusions

QUIS induced SCI resulted in inhibition of GSK-3β in primary afferents and enhanced at-level DRG intrinsic growth (neurite elongation and initiation). Early PI3K mediated activation of GSK-3β attenuated QUIS-induced DRG neurite outgrowth and prevented the development of at-level dysesthesias.

【 授权许可】

   
2015 Bareiss et al.

【 图 表 】

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【 参考文献 】

• [1]Siddall PJ, Loeser JD: Pain following spinal cord injury. Spinal Cord 2001, 39:63-73.
• [2]Widerstrom-Noga E, Biering-Sorensen F, Bryce T, Cardenas DD, Finnerup NB, Jensen MP, et al.: The international spinal cord injury pain basic data set. Spinal Cord 2008, 46:818-823.
• [3]Finnerup NB, Johannesen IL, Sindrup SH, Bach FW, Jensen TS: Pain and dysesthesia in patients with spinal cord injury: a postal survey. Spinal Cord 2001, 39:256-262.
• [4]Siddall PJ, McClelland JM, Rutkowski SB, Cousins MJ: A longitudinal study of the prevalence and characteristics of pain in the first 5 years following spinal cord injury. Pain 2003, 103:249-257.
• [5]Finnerup NB, Norrbrink C, Trok K, Piehl F, Johannesen IL, Sorensen JC, et al.: Phenotypes and predictors of pain following traumatic spinal cord injury: a prospective study. J Pain 2014, 15:40-48.
• [6]Bareiss SK, Gwaltney M, Hernandez K, Lee T, Brewer KL: Excitotoxic spinal cord injury induced dysesthesias are associated with enhanced intrinsic growth of sensory neurons. Neurosci Lett 2013, 542:113-117.
• [7]Bedi SS, Yang Q, Crook RJ, Du J, Wu Z, Fishman HM, et al.: Chronic spontaneous activity generated in the somata of primary nociceptors is associated with pain-related behavior after spinal cord injury. J Neurosci 2010, 30:14870-14882.
• [8]Bedi SS, Lago MT, Masha LI, Crook RJ, Grill RJ, Walters ET: Spinal cord injury triggers an intrinsic growth-promoting state in nociceptors. J Neurotrauma 2012, 29:925-935.
• [9]Yang Q, Wu Z, Hadden JK, Odem MA, Zuo Y, Crook RJ, et al.: Persistent pain after spinal cord injury is maintained by primary afferent activity. J Neurosci 2014, 34:10765-10769.
• [10]Weaver LC, Verghese P, Bruce JC, Fehlings MG, Krenz NR, Marsh DR: Autonomic dysreflexia and primary afferent sprouting after clip-compression injury of the rat spinal cord. J Neurotrauma 2001, 18:1107-1119.
• [11]Christensen MD, Hulsebosch CE: Spinal cord injury and anti-ngf treatment results in changes in cgrp density and distribution in the dorsal horn in the rat. Exp Neurol 1997, 147:463-475.
• [12]Helgren ME, Goldberger ME: The recovery of postural reflexes and locomotion following low thoracic hemisection in adult cats involves compensation by undamaged primary afferent pathways. Exp Neurol 1993, 123:17-34.
• [13]Krenz NR, Meakin SO, Krassioukov AV, Weaver LC: Neutralizing intraspinal nerve growth factor blocks autonomic dysreflexia caused by spinal cord injury. J Neurosci 1999, 19:7405-7414.
• [14]Ondarza AB, Ye Z, Hulsebosch CE: Direct evidence of primary afferent sprouting in distant segments following spinal cord injury in the rat: colocalization of gap-43 and cgrp. Exp Neurol 2003, 184:373-380.
• [15]Krenz NR, Weaver LC: Sprouting of primary afferent fibers after spinal cord transection in the rat. Neuroscience 1998, 85:443-458.
• [16]Woodgett JR: Judging a protein by more than its name: Gsk-3. Sci STKE 2001, 2001:re12.
• [17]Leroy K, Brion JP: Developmental expression and localization of glycogen synthase kinase-3beta in rat brain. J Chem Neuroanat 1999, 16:279-293.
• [18]Kaidanovich-Beilin O, Woodgett JR: Gsk-3: functional insights from cell biology and animal models. Front Mol Neurosci 2011, 4:40.
• [19]Grimes CA, Jope RS: The multifaceted roles of glycogen synthase kinase 3beta in cellular signaling. Prog Neurobiol 2001, 65:391-426.
• [20]Zhou FQ, Zhou J, Dedhar S, Wu YH, Snider WD: NGF-induced axon growth is mediated by localized inactivation of gsk-3beta and functions of the microtubule plus end binding protein apc. Neuron 2004, 42:897-912.
• [21]Rui Y, Myers KR, Yu K, Wise A, De Blas AL, Hartzell HC, et al.: Activity-dependent regulation of dendritic growth and maintenance by glycogen synthase kinase 3beta. Nat Commun 2013, 4:2628.
• [22]Seira O, Del Rio JA: Glycogen synthase kinase 3 beta (gsk3beta) at the tip of neuronal development and regeneration. Mol Neurobiol 2014, 49:931-944.
• [23]Chadborn NH, Ahmed AI, Holt MR, Prinjha R, Dunn GA, Jones GE, et al.: Pten couples sema3a signalling to growth cone collapse. J Cell Sci 2006, 119:951-957.
• [24]Jiang H, Guo W, Liang X, Rao Y: Both the establishment and the maintenance of neuronal polarity require active mechanisms: critical roles of gsk-3beta and its upstream regulators. Cell 2005, 120:123-135.
• [25]Dill J, Wang H, Zhou F, Li S: Inactivation of glycogen synthase kinase 3 promotes axonal growth and recovery in the CNS. J Neurosci 2008, 28:8914-8928.
• [26]Arevalo JC, Chao MV: Axonal growth: where neurotrophins meet Wnts. Curr Opin Cell Biol 2005, 17:112-115.
• [27]Kim YT, Hur EM, Snider WD, Zhou FQ: Role of GSK3 signaling in neuronal morphogenesis. Front Mol Neurosci 2011, 4:48.
• [28]Bareiss S, Kim K, Lu Q: Delta-catenin/nprap: a new member of the glycogen synthase kinase-3beta signaling complex that promotes beta-catenin turnover in neurons. J Neurosci Res 2010, 88:2350-2363.
• [29]Yoshimura T, Kawano Y, Arimura N, Kawabata S, Kikuchi A, Kaibuchi K: Gsk-3beta regulates phosphorylation of crmp-2 and neuronal polarity. Cell 2005, 120:137-149.
• [30]Oh M, Kim H, Yang I, Park JH, Cong WT, Baek MC, et al.: Gsk-3 phosphorylates delta-catenin and negatively regulates its stability via ubiquitination/proteosome-mediated proteolysis. J Biol Chem 2009, 284:28579-28589.
• [31]Wang T, Wu X, Yin C, Klebe D, Zhang JH, Qin X: Crmp-2 is involved in axon growth inhibition induced by rgma in vitro and in vivo. Mol Neurobiol 2013, 47:903-913.
• [32]Alabed YZ, Pool M, Ong Tone S, Sutherland C, Fournier AE: Gsk3 beta regulates myelin-dependent axon outgrowth inhibition through crmp4. J Neurosci 2010, 30:5635-5643.
• [33]Hur EM, Saijilafu , Lee BD, Kim SJ, Xu WL, Zhou FQ: Gsk3 controls axon growth via clasp-mediated regulation of growth cone microtubules. Genes Dev 2011, 25:1968-1981.
• [34]Zhang YK, Huang ZJ, Liu S, Liu YP, Song AA, Song XJ: Wnt signaling underlies the pathogenesis of neuropathic pain in rodents. J Clin Invest 2013, 123:2268-2286.
• [35]Gwak YS, Nam TS, Paik KS, Hulsebosch CE, Leem JW: Attenuation of mechanical hyperalgesia following spinal cord injury by administration of antibodies to nerve growth factor in the rat. Neurosci Lett 2003, 336:117-120.
• [36]Xu JT, Tu HY, Xin WJ, Liu XG, Zhang GH, Zhai CH: Activation of phosphatidylinositol 3-kinase and protein kinase b/akt in dorsal root ganglia and spinal cord contributes to the neuropathic pain induced by spinal nerve ligation in rats. Exp Neurol 2007, 206:269-279.
• [37]Guan XH, Lu XF, Zhang HX, Wu JR, Yuan Y, Bao Q, et al.: Phosphatidylinositol 3-kinase mediates pain behaviors induced by activation of peripheral ephrinbs/ephbs signaling in mice. Pharmacol Biochem Behav 2010, 95:315-324.
• [38]Pezet S, McMahon SB: Neurotrophins: mediators and modulators of pain. Annu Rev Neurosci 2006, 29:507-538.
• [39]Kim WY, Zhou FQ, Zhou J, Yokota Y, Wang YM, Yoshimura T, et al.: Essential roles for gsk-3s and gsk-3-primed substrates in neurotrophin-induced and hippocampal axon growth. Neuron 2006, 52:981-996.
• [40]Hollis ER 2nd, Zou Y: Expression of the wnt signaling system in central nervous system axon guidance and regeneration. Front Mol Neurosci 2012, 5:5.
• [41]Jin Y, Sui HJ, Dong Y, Ding Q, Qu WH, Yu SX, et al.: Atorvastatin enhances neurite outgrowth in cortical neurons in vitro via up-regulating the akt/mtor and akt/gsk-3beta signaling pathways. Acta Pharmacol Sin 2012, 33:861-872.
• [42]Saijilafu , Hur EM, Liu CM, Jiao Z, Xu WL, Zhou FQ: PI3K-GSK3 signalling regulates mammalian axon regeneration by inducing the expression of smad1. Nat Commun. 2013, 4:2690.
• [43]Pezet S, Marchand F, D’Mello R, Grist J, Clark AK, Malcangio M, et al.: Phosphatidylinositol 3-kinase is a key mediator of central sensitization in painful inflammatory conditions. J Neurosci 2008, 28:4261-4270.
• [44]Zhuang ZY, Xu H, Clapham DE, Ji RR: Phosphatidylinositol 3-kinase activates erk in primary sensory neurons and mediates inflammatory heat hyperalgesia through trpv1 sensitization. J Neurosci 2004, 24:8300-8309.
• [45]Xu B, Guan XH, Yu JX, Lv J, Zhang HX, Fu QC, et al.: Activation of spinal phosphatidylinositol 3-kinase/protein kinase b mediates pain behavior induced by plantar incision in mice. Exp Neurol 2014, 255:71-82.
• [46]Yu LN, Zhou XL, Yu J, Huang H, Jiang LS, Zhang FJ, et al.: PI3K contributed to modulation of spinal nociceptive information related to ephrinbs/ephbs. PLoS One 2012, 7:e40930.
• [47]Wang QM, Fiol CJ, DePaoli-Roach AA, Roach PJ: Glycogen synthase kinase-3 beta is a dual specificity kinase differentially regulated by tyrosine and serine/threonine phosphorylation. J Biol Chem 1994, 269:14566-14574.
• [48]Yezierski RP, Liu S, Ruenes GL, Kajander KJ, Brewer KL: Excitotoxic spinal cord injury: behavioral and morphological characteristics of a central pain model. Pain 1998, 75:141-155.
• [49]Brewer KL, Lee JW, Downs H, Oaklander AL, Yezierski RP: Dermatomal scratching after intramedullary quisqualate injection: correlation with cutaneous denervation. J Pain 2008, 9:999-1005.
• [50]Gorman AL, Yu CG, Ruenes GR, Daniels L, Yezierski RP: Conditions affecting the onset, severity, and progression of a spontaneous pain-like behavior after excitotoxic spinal cord injury. J Pain 2001, 2:229-240.
• [51]Yezierski RP: Spinal cord injury pain: spinal and supraspinal mechanisms. J Rehabil Res Dev 2009, 46:95-107.
• [52]Deumens R, Joosten EA, Waxman SG, Hains BC: Locomotor dysfunction and pain: the scylla and charybdis of fiber sprouting after spinal cord injury. Mol Neurobiol 2008, 37:52-63.
• [53]Brown A, Weaver LC: The dark side of neuroplasticity. Exp Neurol 2012, 235:133-141.
• [54]Wong ST, Atkinson BA, Weaver LC: Confocal microscopic analysis reveals sprouting of primary afferent fibres in rat dorsal horn after spinal cord injury. Neurosci Lett 2000, 296:65-68.
• [55]Weaver LC, Cassam AK, Krassioukov AV, Llewellyn-Smith IJ: Changes in immunoreactivity for growth associated protein-43 suggest reorganization of synapses on spinal sympathetic neurons after cord transection. Neuroscience 1997, 81:535-551.
• [56]Polistina DC, Murray M, Goldberger ME: Plasticity of dorsal root and descending serotoninergic projections after partial deafferentation of the adult rat spinal cord. J Comp Neurol 1990, 299:349-363.
• [57]Cole AR, Knebel A, Morrice NA, Robertson LA, Irving AJ, Connolly CN, et al.: Gsk-3 phosphorylation of the alzheimer epitope within collapsin response mediator proteins regulates axon elongation in primary neurons. J Biol Chem 2004, 279:50176-50180.
• [58]Wittmann T, Waterman-Storer CM: Spatial regulation of clasp affinity for microtubules by rac1 and gsk3beta in migrating epithelial cells. J Cell Biol 2005, 169:929-939.
• [59]Watanabe T, Noritake J, Kakeno M, Matsui T, Harada T, Wang S, et al.: Phosphorylation of clasp2 by gsk-3beta regulates its interaction with iqgap1, eb1 and microtubules. J Cell Sci 2009, 122:2969-2979.
• [60]Lucas FR, Goold RG, Gordon-Weeks PR, Salinas PC: Inhibition of gsk-3beta leading to the loss of phosphorylated map-1b is an early event in axonal remodelling induced by wnt-7a or lithium. J Cell Sci 1998, 111:1351-1361.
• [61]Sanchez C, Diaz-Nido J, Avila J: Phosphorylation of microtubule-associated protein 2 (map2) and its relevance for the regulation of the neuronal cytoskeleton function. Prog Neurobiol 2000, 61:133-168.
• [62]Trivedi N, Marsh P, Goold RG, Wood-Kaczmar A, Gordon-Weeks PR: Glycogen synthase kinase-3beta phosphorylation of map1b at ser1260 and thr1265 is spatially restricted to growing axons. J Cell Sci 2005, 118:993-1005.
• [63]Garrido JJ, Simon D, Varea O, Wandosell F: Gsk3 alpha and gsk3 beta are necessary for axon formation. FEBS Lett 2007, 581:1579-1586.
• [64]Li CL, Sathyamurthy A, Oldenborg A, Tank D, Ramanan N: Srf phosphorylation by glycogen synthase kinase-3 promotes axon growth in hippocampal neurons. J Neurosci 2014, 34:4027-4042.
• [65]Zhao T, Qi Y, Li Y, Xu K: Pi3 kinase regulation of neural regeneration and muscle hypertrophy after spinal cord injury. Mol Biol Rep 2012, 39:3541-3547.
• [66]Ma W, Chabot JG, Quirion R: A role for adrenomedullin as a pain-related peptide in the rat. Proc Natl Acad Sci USA 2006, 103:16027-16032.
• [67]Brown A, Ricci MJ, Weaver LC: Ngf message and protein distribution in the injured rat spinal cord. Exp Neurol 2004, 188:115-127.
• [68]Brown A, Ricci MJ, Weaver LC: NGF mRNA is expressed in the dorsal root ganglia after spinal cord injury in the rat. Exp Neurol 2007, 205:283-286.
• [69]Zhou XF, Deng YS, Xian CJ, Zhong JH: Neurotrophins from dorsal root ganglia trigger allodynia after spinal nerve injury in rats. Eur J Neurosci 2000, 12:100-105.
• [70]Itokazu T, Hayano Y, Takahashi R, Yamashita T: Involvement of wnt/beta-catenin signaling in the development of neuropathic pain. Neurosci Res 2014, 79:34-40.
• [71]Krenz NR, Weaver LC: Nerve growth factor in glia and inflammatory cells of the injured rat spinal cord. J Neurochem 2000, 74:730-739.
• [72]Christensen MD, Hulsebosch CE: Chronic central pain after spinal cord injury. J Neurotrauma 1997, 14:517-537.
• [73]Chao MV: Neurotrophins and their receptors: a convergence point for many signalling pathways. Nat Rev Neurosci 2003, 4:299-309.
• [74]Yoshimura T, Arimura N, Kaibuchi K: Molecular mechanisms of axon specification and neuronal disorders. Ann NY Acad Sci 2006, 1086:116-125.
• [75]Cosker KE, Eickholt BJ: Phosphoinositide 3-kinase signalling events controlling axonal morphogenesis. Biochem Soc Trans 2007, 35:207-210.
• [76]Yoshimura T, Arimura N, Kaibuchi K: Signaling networks in neuronal polarization. J Neurosci 2006, 26:10626-10630.
• [77]Zhou FQ, Snider WD: Intracellular control of developmental and regenerative axon growth. Philos Trans R Soc Lond B Biol Sci 2006, 361:1575-1592.
• [78]Markus A, Zhong J, Snider WD: Raf and akt mediate distinct aspects of sensory axon growth. Neuron 2002, 35:65-76.
• [79]Eickholt BJ, Walsh FS, Doherty P: An inactive pool of gsk-3 at the leading edge of growth cones is implicated in semaphorin 3a signaling. J Cell Biol 2002, 157:211-217.
• [80]Weng HR, Gao M, Maixner DW: Glycogen synthase kinase 3 beta regulates glial glutamate transporter protein expression in the spinal dorsal horn in rats with neuropathic pain. Exp Neurol 2014, 252:18-27.
• [81]Shi TJ, Huang P, Mulder J, Ceccatelli S, Hokfelt T: Expression of p-akt in sensory neurons and spinal cord after peripheral nerve injury. Neurosignals 2009, 17:203-212.
• [82]Jope RS, Yuskaitis CJ, Beurel E: Glycogen synthase kinase-3 (gsk3): inflammation, diseases, and therapeutics. Neurochem Res 2007, 32:577-595.
• [83]Cuzzocrea S, Genovese T, Mazzon E, Crisafulli C, Di Paola R, Muia C, et al.: Glycogen synthase kinase-3 beta inhibition reduces secondary damage in experimental spinal cord trauma. J Pharmacol Exp Ther 2006, 318:79-89.
• [84]Martins DF, Rosa AO, Gadotti VM, Mazzardo-Martins L, Nascimento FP, Egea J, et al.: The antinociceptive effects of ar-a014418, a selective inhibitor of glycogen synthase kinase-3 beta, in mice. J Pain 2011, 12:315-322.
• [85]Mazzardo-Martins L, Martins DF, Stramosk J, Cidral-Filho FJ, Santos AR: Glycogen synthase kinase 3-specific inhibitor ar-a014418 decreases neuropathic pain in mice: evidence for the mechanisms of action. Neuroscience 2012, 226:411-420.
• [86]Tuncdemir M, Yildirim A, Karaoglan A, Akdemir O, Ozturk M: Ar-a014418 as a glycogen synthase kinase-3 inhibitor: anti-apoptotic and therapeutic potential in experimental spinal cord injury. Neurocirugia 2013, 24:22-32.
• [87]Cyranoski D: Chinese network to start trials of spinal surgery. Nature 2007, 446:476-477.
• [88]Yick LW, So KF, Cheung PT, Wu WT: Lithium chloride reinforces the regeneration-promoting effect of chondroitinase abc on rubrospinal neurons after spinal cord injury. J Neurotrauma 2004, 21:932-943.
• [89]Wang Y, Gu Q, Dong Y, Zhou W, Song H, Liu Y, et al.: Inhibition of gecko gsk-3beta promotes elongation of neurites and oligodendrocyte processes but decreases the proliferation of blastemal cells. J Cell Biochem 2012, 113:1842-1851.
• [90]Sapunar D, Kostic S, Banozic A, Puljak L: Dorsal root ganglion—a potential new therapeutic target for neuropathic pain. J Pain Res 2012, 5:31-38.
• [91]Krames ES: The role of the dorsal root ganglion in the development of neuropathic pain. Pain Med 2014, 10:1669-1685.
• [92]Carlton SM, Du J, Tan HY, Nesic O, Hargett GL, Bopp AC, et al.: Peripheral and central sensitization in remote spinal cord regions contribute to central neuropathic pain after spinal cord injury. Pain 2009, 147:265-276.
• [93]Yezierski RP, Santana M, Park SH, Madsen PW: Neuronal degeneration and spinal cavitation following intraspinal injections of quisqualic acid in the rat. J Neurotrauma 1993, 10:445-456.
• [94]Twiss JL, Smith DS, Chang B, Shooter EM: Translational control of ribosomal protein l4 mrna is required for rapid neurite regeneration. Neurobiol Dis 2000, 7:416-428.
• [95]Kalous A, Osborne PB, Keast JR: Spinal cord compression injury in adult rats initiates changes in dorsal horn remodeling that may correlate with development of neuropathic pain. J Comp Neurol 2009, 513:668-684.
• [96]Yezierski RP, Park SH: The mechanosensitivity of spinal sensory neurons following intraspinal injections of quisqualic acid in the rat. Neurosci Lett 1993, 157:115-119.
• [97]Liu S, Ruenes GL, Yezierski RP: NMDA and non-NMDA receptor antagonists protect against excitotoxic injury in the rat spinal cord. Brain Res 1997, 756:160-167.